Impact of exercise on brain-bone marrow interactions in chronic stress: potential mechanisms preventing stress-induced hypertension.

We examined the effect of chronic restraint stress and the counteractive effects of daily exercise on the molecular basis of the brain-bone marrow (BM) interactions, by especially focusing on the paraventricular nucleus (PVN) of the hypothalamus. Male Wistar rats were assigned into control, restraint stress, and stress + daily spontaneous exercise (SE) groups. BM and hypothalamic gene expression profiles were examined through the undertaking of RT-PCR and microarrays, respectively. The inflammatory blood cell population was investigated through flow cytometry. Through the use of immunohistochemistry, we examined the presence of BM-derived C-C chemokine receptor type 2 (CCR2)-expressing microglial cells in the rat PVN. The gene expression levels of BM inflammatory factors such as those of interleukin 1 beta and CCR2, and the inflammatory blood cell population were found to be significantly higher in both restrained groups compared with control group. Interestingly, chronic restraint stress alone activated the recruitment of BM-derived CCR2-expressing microglial cells into the PVN, whereas daily spontaneous exercise prevented it. A notable finding was that restraint stress upregulated relative gene expression of hypothalamic matrix metalloproteinase 3 (MMP3), which increases the permeability of the blood-brain barrier (BBB), and that exercise managed to normalize it. Moreover, relative expression of some hypothalamic genes directly involved in the facilitation of cell migration was downregulated by daily exercise. Our findings suggest that daily spontaneous exercise can reduce the numbers of BM-derived CCR2-expressing microglial cells into the PVN through the prevention of stress-induced changes in the hypothalamic gene expression.NEW & NOTEWORTHY Chronic restraint stress can upregulate MMP3 gene expression in the rat hypothalamus, whereas daily spontaneous exercise can prevent this stress-induced effect. Stress-induced BM-derived inflammatory cell recruitment into the rat PVN can be prevented by daily spontaneous exercise. Stress-induced increase of hypothalamic MMP3 gene expression may be responsible for BBB injury, thereby allowing for BM-derived inflammatory cells to be recruited and to accumulate in the rat PVN, and to be subsequently involved in the onset of stress-induced hypertension.

Iron Induces Resistance Against the Rice Blast Fungus Magnaporthe oryzae Through Potentiation of Immune Responses.

Iron is an essential nutrient required for plant growth and development. The availability of iron might also influence disease resistance in plants. However, the molecular mechanisms involved in the plant response to iron availability and immunity have been investigated separately from each other. In this work, we found that exposure of rice plants to high iron enhances resistance to infection by the fungal pathogen Magnaporthe oryzae, the causal agent of blast disease. RNA-Seq analysis revealed that blast resistance in iron-treated rice plants was associated with superinduction of defense-related genes during pathogen infection, including Pathogenesis-Related genes. The expression level of genes involved in the biosynthesis of phytoalexins, both diterpene phytoalexins and the flavonoid phytoalexin sakuranetin, was also higher in iron-treated plants compared with control plants, which correlated well with increased levels of phytoalexins in these plants during M. oryzae infection. Upon pathogen infection, lipid peroxidation was also higher in iron-treated plants compared with non-treated plants. We also show that M. oryzae infection modulates the expression of genes that play a pivotal role in the maintenance of iron homeostasis. Histochemical analysis of M. oryzae-infected leaves revealed colocalization of iron and reactive oxygen species in cells located in the vicinity of fungal penetration sites (e.g. appressoria) in rice plants that have been exposed to iron. Together these findings support that ferroptosis plays a role in the response of iron-treated rice plants to infection by virulent M. oryzae. Understanding interconnected regulations between iron signaling and immune signaling in rice holds great potential for developing novel strategies to improve blast resistance in rice.

Efficacy of edoxaban for the treatment of gynecological cancer-associated venous thromboembolism: analysis of Japanese real-world data.

OBJECTIVE: Direct oral anticoagulants (DOACs) are increasingly being used for the treatment of cancer-associated venous thromboembolism (CAT). However, there is limited evidence of the efficacy of DOACs for the treatment of gynecological CAT. Thus, this study aimed to investigate the efficacy and safety of edoxaban for the treatment of gynecological CAT using Japanese real-world data. METHODS: We reviewed the medical records of patients with 371 gynecological cancer who received edoxaban or vitamin K antagonist (VKA) between January 2011 and December 2018. RESULTS: Altogether, 211 and 160 patients were treated with edoxaban and VKA, respectively. Fourteen patients (6.8%) in the edoxaban group and 22 (13.8%) in the VKA group showed recurrence of venous thromboembolism (VTE). Cumulative VTE recurrence was not significantly different between the 2 groups (p=0.340). Adverse events occurred in 15 (7.1%) and 11 (6.9%) patients in the edoxaban and VKA groups, respectively (p=0.697). Subgroup analysis of the edoxaban and VKA groups according to different tumor types, including ovarian, endometrial, and cervical cancer, showed equivalent outcomes in terms of VTE recurrence and adverse events. Patients without pulmonary embolism (PE) were mostly omitted from initial unfractionated heparin (UFH) therapy prior to administration of edoxaban. However, this did not increase the recurrence of VTE. CONCLUSION: This study confirmed that edoxaban is effective and safe for the treatment of gynecological CAT. This finding was consistent for different types of gynecological cancer. Additionally, initial UFH therapy prior to the administration of edoxaban may be unnecessary for patients without PE.

Clinical Availability of Tumour Biopsy Using Diagnostic Laparoscopy for Advanced Ovarian Cancer.

BACKGROUND/AIM: Tumour biopsy using laparoscopy before neoadjuvant chemotherapy for advanced ovarian cancer has been widely accepted. However, there are few reports about its operative outcome compared to biopsy with laparotomy. We investigated the advantage of laparoscopic biopsy for advanced ovarian cancer. PATIENTS AND METHODS: We included 23 patients who underwent laparoscopy and 27 who underwent exploratory laparotomy before neoadjuvant chemotherapy between January 2012 and August 2020. We reviewed their medical records and evaluated their operative outcomes. RESULTS: Blood loss was significantly lower in the laparoscopy group (5 ml vs. 320 ml, p<0.05). The period until the initiation of neoadjuvant chemotherapy was significantly shorter in the laparoscopy group (12 days vs. 16 days, p<0.05). Overall survival did not differ significantly between the two groups (25.4 months vs. 24.7 months, p=0.53). CONCLUSION: Laparoscopic tumour biopsy is useful and safe for histological diagnosis, thereby allowing for early introduction to neoadjuvant chemotherapy.

Characterization of microRNA expression in B cells derived from Japanese black cattle naturally infected with bovine leukemia virus by deep sequencing.

Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis (EBL), a malignant B cell lymphoma. However, the mechanisms of BLV-associated lymphomagenesis remain poorly understood. Here, after deep sequencing, we performed comparative analyses of B cell microRNAs (miRNAs) in cattle infected with BLV and those without BLV. In BLV-infected cattle, BLV-derived miRNAs (blv-miRNAs) accounted for 38% of all miRNAs in B cells. Four of these blv-miRNAs (blv-miR-B1-5p, blv-miR-B2-5p, blv-miR-B4-3p, and blv-miR-B5-5p) had highly significant positive correlations with BLV proviral load (PVL). The read counts of 90 host-derived miRNAs (bta-miRNAs) were significantly down-regulated in BLV-infected cattle compared to those in uninfected cattle. Only bta-miR-375 had a positive correlation with PVL in BLV-infected cattle and was highly expressed in the B cell lymphoma tissue of EBL cattle. There were a few bta-miRNAs that correlated with BLV tax/rex gene expression; however, BLV AS1 expression had a significant negative correlation with many of the down-regulated bta-miRNAs that are important for tumor development and/or tumor suppression. These results suggest that BLV promotes lymphomagenesis via AS1 and blv-miRNAs, rather than tax/rex, by down-regulating the expression of bta-miRNAs that have a tumor-suppressing function, and this downregulation is linked to increased PVL.

Analysis of risk factors for patients with early-stage cervical cancer: A study of 374 patients.

AIM: This study aimed to identify the postoperative histological features affecting the prognosis of patients with early-stage cervical cancer who underwent open radical hysterectomy. METHODS: This retrospective study enrolled 374 patients with pT1a, 1b1 and 2a1 early-stage cervical cancer who underwent open radical hysterectomy between 2001 and 2018. Survival outcomes were analyzed by Kaplan-Meier method and compared with log-rank test. Using the Cox proportional hazards regression test, we conducted a multivariate analysis for disease-free survival and overall survival. RESULTS: Others histology, including other epithelial tumors and neuroendocrine tumors, had a significantly worse prognosis in both disease-free survival and overall survival than those of squamous cell carcinoma and adenocarcinoma (hazard ratio, 4.37 and 11.76; P = 0.006 and P = 0.002, respectively), along with lymph node metastasis (hazard ratio, 2.99 and 7.03; P = 0.009 and P = 0.001, respectively). CONCLUSION: Others histology including adenosquamous carcinoma had a poor prognosis in early-stage cervical cancer as with high-risk factors.

Effectiveness of on-farm continuous flow high-temperature short-time pasteurization for inactivation of bovine leukemia virus in milk.

The effectiveness of on-farm continuous flow high-temperature short-time (HTST) pasteurization (i.e., 72°C for 15 s) for the inactivation of bovine leukemia virus (BLV) in milk was investigated with a sheep bioassay. Four sheep that had been inoculated with completely pasteurized milk containing approximately 3.4 × 107 BLV-infected peripheral blood mononuclear cells (PBMC) and treated by either HTST pasteurization or laboratory-scale low-temperature long-time (LTLT) pasteurization (i.e., 60°C for 30 min), remained negative for BLV for at least 17 weeks after inoculation. In contrast, all sheep inoculated with unpasteurized or inadequately pasteurized milk containing the same number of BLV-infected PBMC were tested positive for BLV and anti-BLV antibodies within 3 weeks after inoculation. These results suggest that on-farm continuous flow HTST pasteurization was equivalent value with inactivated BLV on the LTLT procedure and can effectively inactivate BLV in the milk. Therefore, on-farm HTST pasteurization of the pooled colostrum or milk used in automated feeding systems is likely to protect group-housed preweaned calves from BLV infection, thereby improving animal health on dairy farms.

Development of a microchip electrophoresis-based, high-throughput PCR-RFLP method to type Tax 233 variants of bovine leukemia virus in Japan.

Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis (EBL). We used microchip electrophoresis in combination with automatic image analysis to develop a novel high-throughput PCR-RFLP to type the gene sequences that encode BLV Tax 233. This method revealed that 233L-Tax is more prevalent than 233P-Tax in cattle in Japan. The proportion infected with BLV carrying the gene encoding 233L-Tax was significantly higher in Holstein cattle than in Japanese Black cattle. Holsteins infected with BLV encoding 233L-Tax had higher proviral loads than did Holsteins infected with BLV encoding 233P-Tax and Japanese Blacks infected with BLV encoding 233L-Tax or 233P-Tax. The novel method developed in this study will be a useful tool for identifying cattle harboring BLV with a higher risk of EBL and viral transmission.

Genomic evidence for convergent evolution of gene clusters for momilactone biosynthesis in land plants.

Momilactones are bioactive diterpenoids that contribute to plant defense against pathogens and allelopathic interactions between plants. Both cultivated and wild grass species of Oryza and Echinochloa crus-galli (barnyard grass) produce momilactones using a biosynthetic gene cluster (BGC) in their genomes. The bryophyte Calohypnum plumiforme (formerly Hypnum plumaeforme) also produces momilactones, and the bifunctional diterpene cyclase gene CpDTC1/HpDTC1, which is responsible for the production of the diterpene framework, has been characterized. To understand the molecular architecture of the momilactone biosynthetic genes in the moss genome and their evolutionary relationships with other momilactone-producing plants, we sequenced and annotated the C. plumiforme genome. The data revealed a 150-kb genomic region that contains two cytochrome P450 genes, the CpDTC1/HpDTC1 gene and the "dehydrogenase momilactone A synthase" gene tandemly arranged and inductively transcribed following stress exposure. The predicted enzymatic functions in yeast and recombinant assay and the successful pathway reconstitution in Nicotiana benthamiana suggest that it is a functional BGC responsible for momilactone production. Furthermore, in a survey of genomic sequences of a broad range of plant species, we found that momilactone BGC is limited to the two grasses (Oryza and Echinochloa) and C. plumiforme, with no synteny among these genomes. These results indicate that while the gene cluster in C. plumiforme is functionally similar to that in rice and barnyard grass, it is likely a product of convergent evolution. To the best of our knowledge, this report of a BGC for a specialized plant defense metabolite in bryophytes is unique.

Surgical results in older patients with lumbar spinal stenosis according to gait speed in relation to the diagnosis for sarcopenia.

PURPOSE: The purpose of this study was to evaluate the implication of gait speed, which is a prerequisite for the diagnosis of sarcopenia in older patients with lumbar spinal stenosis (LSS). METHODS: This study was conducted in a total of 235 patients with LSS who underwent surgical treatment. The state of sarcopenia and pre-sarcopenia, including gait speed, were evaluated before and after the operation. RESULTS: The proportion of patients with lower than at baseline levels of skeletal muscle mass index (SMI) and gait speed was 27.2% and 17.9%, respectively. Significant changes were observed in gait speed, whereas SMI showed no significant differences between the preoperative and postoperative periods. Sarcopenic patients presented lower levels of activities of daily living preoperatively than pre-sarcopenic patients. However, favorable surgical results were obtained postoperatively. Significant changes were observed in gait speed, whereas grip strength and SMI showed no significant differences between the preoperative and postoperative periods. The postoperative decrease in SMI was not significant. CONCLUSION: Postoperative gait speed was significantly improved, whereas muscle mass did not increase in the patients in this study. Therefore, low gait speed in patients with LSS seems to be derived from a neurologic disorder. The surgical results in low muscle mass patients without low gait speed were similar to those with low gait speed. Sarcopenia in elderly patients with locomotor disease should be evaluated using muscle mass alone without assessing physical performance.

Surgical Treatment Assessment of Cervical Laminoplasty Using Quantitative Performance Evaluation in Elderly Patients: A Prospective Comparative Study in 505 Patients With Cervical Spondylotic Myelopathy.

STUDY DESIGN: A prospective cohort study. OBJECTIVE: The purpose of this study was to compare surgical outcomes between non-elderly and elderly patients with cervical spondylotic myelopathy (CSM) who underwent laminoplasty. SUMMARY OF BACKGROUND DATA: Since age at the time of surgery influences the surgical outcome, we designed a large-scale cohort study to examine the surgical outcome for CSM from a single operative procedure used exclusively in elderly patients. METHODS: A total of 505 consecutive patients with CSM (311 men; 194 women) were prospectively enrolled. The mean age was 66.6 years (range, 41-91), and the average postoperative follow-up period was 26.5 ±â€Š12.5 months. Patients were divided into three groups according to age: non-elderly (<65 yr, n = 201), young-old (65-74 yr, n = 186), and old-old (≥75 yr, n = 118). Pre- and postoperative neurological status was evaluated using the Japanese Orthopaedic Association scoring system for cervical myelopathy (JOA score) and quantifiable tests-the 10-s grip and release test (10-s G&R test) and the 10-s step test. RESULTS: Mean achieved JOA scores in non-elderly, young-old, and old-old groups were 3.1, 3.2, and 3.0, respectively, with no significant difference among three groups (P = 0.5735). Mean preoperative 10-s G&R test results were 17.3, 14.4, and 13.0, respectively, indicating a significant decrease with increasing age, whereas postoperative results significantly improved in all groups (21.0, 17.9, and 16.3, respectively). Similarly, the 10-s step test significantly decreased with age, with preoperative scores of 14.3, 11.5, and 8.6, respectively, whereas postoperative scores improved to 17.3, 14.9, and 12.5, respectively. The three groups showed no significant difference in the rate of postoperative complications. CONCLUSION: Elderly patients adequately recovered from laminoplasty in terms of achieved JOA score, the 10-s G&R test, and the 10-s step test. Therefore, laminoplasty for CSM is beneficial in elderly patients. LEVEL OF EVIDENCE: 2.

Septic shock due to infected giant venous malformation complicated by massive bleeding.

Case: A 19-year-old man with a hemangioma that extended from the left arm to the axillary region had deteriorated due to shock, and no improvement was observed after fluid resuscitation. His status on arrival led to complications of hemorrhagic and septic shock with his left arm swollen and deep purple in color. Left arm amputation to control the source of bleeding and infection was thought to be indicated, however, the coagulation system had collapsed. Therefore, initial reduction of the blood flow to the hemangioma with angiographic endovascular treatment was carried out. Thereafter, continuous hemodiafiltration using a polymethylmethacrylate membrane hemofilter against hypercytokinemia was introduced in the intensive care unit for damage control, which resulted in success. Outcome: We completed the arm amputation, and subsequently carried out a latissimus dorsi muscle flap transfer on the amputated stump. The patient achieved an ambulatory discharge. Conclusions: We successfully treated the very rare case of massive venous malformation with shock due to hemorrhage and infection by performing damage control.

Treatment outcomes of intradiscal steroid injection/selective nerve root block for 161 patients with cervical radiculopathy.

Patients with cervical radiculopathy (CR) were treated with intradiscal injection of steroids (IDIS) and/or selective nerve root block (SNRB) at our hospital. We retrospectively report the outcomes of these nonsurgical treatments for CR. 161 patients who were followed up for >2months were enrolled in this study. Patients' clinical manifestations were classified as arm pain, arm numbness, neck and/or scapular pain, and arm paralysis. Improvement in each manifestation was classified as "disappeared," "improved," "poor," or "worsened." Responses of "disappeared" or "improved" manifestations suggested treatment effectiveness. Final clinical outcomes were evaluated using the Odom criteria. Changes in herniated disc size were evaluated by comparing the initial and final MRI scans. On the basis of these changes, the patients were divided into regression, no-change, or progression groups. We investigated the relationship between the Odom criteria and changes observed on MRI. Effectiveness rates were 89% for arm pain, 77% for arm numbness, 82% for neck and/or scapular pain, and 76% for arm paralysis. In total, 91 patients underwent repeated MRI. In 56 patients (62%), the size of the herniated disc decreased, but 31 patients (34%) exhibited no change in disc size. The regression group showed significantly better Odom criteria results than the no-change group. In conclusion, IDIS and SNRB for CR are not widely performed. However, other extremely effective therapies that can rapidly improve neuralgia should be considered before surgery.

Functional computed tomography scanning for evaluating fusion status after anterior cervical decompression fusion.

INTRODUCTION: Nonunion is a major complication of anterior cervical fusion that causes poor outcomes and occasionally requires additional operative intervention. The purpose of this study is to evaluate the accuracy of functional computed tomography (CT) scanning for determining fusion status after anterior cervical fusion by comparing with functional radiographs. MATERIALS AND METHODS: The fusion status in 59 patients treated by anterior cervical fusion was assessed by functional radiography and functional CT scanning at 6 and 12 months after surgery. Fusion rates and clinical symptoms were evaluated. Fusion on functional radiography was defined as less than 2 mm of motion between adjacent spinous processes and a particular bony trabeculation on functional CT; fusion was defined as nonexistence of a clear zone or a gas pattern and a particular bone connection on reconstructed sagittal-view images. RESULTS: Functional radiographs demonstrated solid fusion in 83.9% at 6 months and 91.1% at 12 months postoperatively; functional CT showed solid fusion in 55.3 and 78.6%, respectively. The fusion rate detected on functional CT images was significantly lower than that on functional radiographs at each period. At 6 months postoperatively, patients with incomplete union on functional CT were more likely to have neck pain than those who had complete union on functional CT. (46.2 vs 13.3%, P < 0.05) CONCLUSION: Functional CT can detect nonunion more clearly than functional radiography. At 6 months postoperatively, patients with incomplete union on functional CT images were likely to have more neck pain. Functional CT may allow accurate detection of symptomatic nonunion after anterior cervical fusion.

Risk factors for poor outcome of cervical laminoplasty for cervical spondylotic myelopathy in patients with diabetes.

BACKGROUND: Diabetes is one of the most frequent comorbidities in patients with cervical spondylotic myelopathy. The purpose of this study was to characterize the risk factors for poor treatment outcome following cervical laminoplasty for cervical spondylotic myelopathy in patients with diabetes. METHODS: A total of 105 consecutive patients with diabetes and cervical spondylotic myelopathy who underwent double-door laminoplasty were included in this study; there were sixty-five male patients and forty female patients with a mean age of 68.2 years (range, forty-three to eighty-three years). All patients were followed for twelve months or more after surgery, with a mean follow-up time (and standard deviation) of 25.7 ± 14.2 months. All patients had palliative controlled blood glucose levels in the immediate term during the perioperative period. We evaluated the recovery rate on the basis of the Japanese Orthopaedic Association score. Logistic regression analyses (univariate analysis and multivariate analysis) were performed to identify the risk factors for poor outcome. Poor outcome was defined as a postoperative recovery rate of <50%, and good outcome was defined as a postoperative recovery rate of ≥ 50%. RESULTS: Univariate logistic regression analysis showed that a patient age of sixty-five years or older (odds ratio, 3.111; p = 0.0085) and a duration of cervical spondylotic myelopathy symptoms for twelve months or more (odds ratio, 3.940; p = 0.0012) were associated with an increased risk of poor surgical outcome. High glycated hemoglobin levels of ≥ 6.5% (odds ratio, 2.591; p = 0.0193) and a duration of diabetes for ten years or more (odds ratio, 2.245; p = 0.0321) were significant risk factors for poor surgical outcome. Multivariate logistic regression analysis determined that glycated hemoglobin levels of ≥ 6.5% (odds ratio, 2.822; p = 0.0441) and a duration of diabetes for ten years or more (odds ratio, 2.240; p = 0.0410) were significant risk factors for poor treatment outcome. Fasting blood glucose levels did not affect treatment outcomes. CONCLUSIONS: Diabetes with advanced age and long-term cervical spondylotic myelopathy symptoms adversely affected cervical laminoplasty outcomes. High preoperative glycated hemoglobin levels and long-term diabetes are risk factors for poor cervical laminoplasty outcomes in patients with diabetes and cervical spondylotic myelopathy.

Impact of diabetes on the outcomes of cervical laminoplasty: a prospective cohort study of more than 500 patients with cervical spondylotic myelopathy.

STUDY DESIGN: A prospective cohort study. OBJECTIVE: This study aimed to compare the outcome of cervical laminoplasty between diabetic patients and nondiabetic patients with cervical spondylotic myelopathy. SUMMARY OF BACKGROUND DATA: A few retrospective studies have investigated the outcomes of cervical myelopathy in diabetic patients; therefore, our large-scale cohort study was designed to assess these outcomes in cervical spondylotic myelopathy. METHODS: In total, 505 consecutive patients with cervical spondylotic myelopathy (311 males, 194 females; mean age, 66.6 yr; range, 41-91 yr) who underwent double-door laminoplasty were prospectively enrolled. They were followed up for more than 12 months after surgery (mean follow-up period, 25.6 ± 12.6 mo). The patients were divided on the basis of diabetic criteria for glucose intolerance into 2 groups: the diabetic group (n = 105) and nondiabetic group (n = 400). We evaluated differences in pre- and postoperative Japanese Orthopaedic Association (JOA) scores, recovery rate, achieved JOA scores (postoperative JOA score - preoperative JOA score), and complications between both groups. RESULTS: The mean JOA scores in the diabetic and nondiabetic groups were 10.1 and 10.8 points before surgery and 13.1 and 13.9 points after surgery, respectively. The diabetic group showed significantly low pre- and postoperative JOA scores and low recovery rate of JOA scores compared with the nondiabetic group (47.3% vs. 53.6%, P < 0.05). However, mean achieved JOA scores in the diabetic and nondiabetic groups were 3.0 and 3.1 points respectively, with no significant difference between both groups (P = 0.343). The groups showed no significant difference in the postoperative complication rate. CONCLUSION: Pre- and postoperative JOA scores and recovery rates were lower in the diabetic group than the nondiabetic group. However, the achieved JOA scores were not significantly different between both groups. Diabetic and nondiabetic patients experienced similar benefits from laminoplasty. LEVEL OF EVIDENCE: 3.

A novel mutation of the bovine EDA gene associated with anhidrotic ectodermal dysplasia in Holstein cattle.

Anhidrotic ectodermal dysplasia (EDA) is a genetic disease characterized by the absence or hypoplasia of hair, teeth and eccrine sweat glands that has been reported in humans, the tabby mouse mutants, cattle and dogs. The EDA gene on the X chromosome encodes a protein, ectodysplasin-A (EDA), which is responsible for EDA. Here we describe a novel mutation of the EDA gene in which a 19 bp deletion in exon 1 in male Holstein calves demonstrated the phenotypic features of EDA. The dam and the grand-dam of the affected calves were heterozygous for this deletion. It is assumed that this deletion close to the start codon confuses all transcripts, and leads to the complete loss of pleiotropic functions of the bovine EDA gene. These results suggest that this mutation might be useful as animal models for the investigation of the pathogenic mechanisms of the anhidrotic ectodermal dysplasia.